Plasmids |
pALS297 (ttTi5605-MCS loxP::Prps-27::NeoR::unc-54-3ʹutr::loxP); Addgene # 129538 |
pALS186 (ttTi5605-MCS loxP::C.br.unc-119(+)::loxP); Addgene # 129539 |
Strains |
EG6699 ttTi5605(mos1) II; unc-119(ed3), CGC, note lost extrachromosomal array oxEx1578 |
ALS344 ttTi5605(mos1) II, this study; EG6699 was crossed 4x to N2 to remove the unc-119(ed3) allele |
Primers to follow ttTi5605(mos1) II |
o508-mos1F(oJL103) TCTGCGAGTTGTTTTTGCGTTTGAG |
o509-ttTi5605F GATTGTTTGACCTGGCGGAACT |
o91-ttTi55605R(NM3888) ACGCCCAGGAGAACACGTTAG |
Primers for cloning |
op297-p186AvrIIF TAGAGGGTACCAGAGCTCACCTAGGN*20-22 |
op297XhoIR GTTATaCGCGCACCGTACGTCTCGAGN*20-22 |
op186XhoIR GTTATgCGCGCACCGTACGTCTCGAGN*20-22 |
Primers for insert verification |
5ʹarm, neoR mosSCI |
o145-ttTi5605outerF AGGCAGAATGTGAACAAGACTCGAG (NM3880 + 2nt) |
o496-unc54-3ʹutrR GGCCCAGACGTGCGAAGAAATA (or transgene-specific REV primer for long transgenes) |
3ʹarm, unc-119 mosSCI |
o145-ttTi5605outerF AGGCAGAATGTGAACAAGACTCGAG (NM3880 + 2nt) |
o147-Cbr-unc-119promR AAGTAGCAGAGCTGGGGAGAAGAA (or transgene-specific REV primer for long transgenes) |
5ʹarm, neoR mosSCI |
o311-NeoRF CTTCTATCGCCTTCTTGACGAG |
o146-ttTi5605outerR AATCGGGAGGCGAACCTAACTGT (NM3884 + 2nt) |
3ʹarm, unc-119 mosSCI |
Transgene-specific FOR primer |
o146-ttTi5605outerR AATCGGGAGGCGAACCTAACTGT (NM3884 + 2nt) |